同种异体骨髓基质细胞肌肉内成骨分化的实验研究

目的 观察并探讨同种异体兔骨髓基质细胞在未接受免疫抑制剂的正常兔肌肉内移植后的免疫原性、存活及成骨状况.方法 首先通过兔谱系差异选择供、受体,并采用淋巴细胞反应进一步证实免疫学错配,之后抽取兔骨髓、体外分离培养、获取骨髓基质细胞,体外加矿化条件培养液诱导成骨、并对成骨特性予以鉴定后,将异体骨髓基质细胞移植入受体兔肌肉内,从淋巴细胞转化率、淋巴细胞杀伤活性、组织学检查、种子细胞体内成活及成骨情况等方面检测. 结果 术后各检测时间,异体骨髓基质细胞移植未诱发淋巴细胞大量增殖反应(P>0.05),淋巴细胞杀伤活性与自体组差异无统计学意义(P>0.05),移植区未见到大量炎性细胞浸润,经组织学及免疫组化染色证实异体骨髓基质细胞在受体组织内存活,8周后于异体肌肉中形成骨组织. 结论 同种异体骨髓基质细胞移植,未诱发免疫排斥反应,在无免疫抑制剂应用的情况下,移植细胞在受体肌肉内存活、并形成骨性组织.

Experimental study on osteogenic differentiation of allogeneic bone marrow stromal cells in muscles of rabbits

Objective To study and evaluate the ability of allogeneic bone marrow stromal cells(BMSCs) to survive and regenerate bone in muscles without using immunosuppressive agents. Methods A complete mismatch between donor BMSCs and recipient rabbit was confirmed by one way mixed lymphocyte reaction assaysprior to implantation. And then bone marrow aspirates were obtained from donor rabbits.BMSCs were isolated from the bone marrow, cultured in conditional medium to be ubdyced to become osteogenic, and then to examine these characteristics. After that the donor BMSCs were transplanted into the recipient rabbits. Immunological tests such as lymphocyte transformation rate and cell mediated cytotoxity, histological observation, seeding cells survival and bone formation were performed following transplantation. Results Allogeneic BMSCs transplantation did not actually elicit an adverse immune response, and bone regenerated at the transplantation area, and then the transplanted BMSCs marked with BRDU preoperatively were founded to be living 8 weeks later after transplantation using immunohistochemistric technique. Conclusion Allogeneic BMSCs would not elicit an adverse immune response in vivo without the immunosuppressive therapy, which could survive and form new bone tissue with the help of BMP-2 in the muscles.